Laylah Perez
Pronouns: she/her
Research Mentor(s): Steven Huang
Research Mentor School/College/Department: Department of Internal Medicine, Division of Pulmonary and Critical Care / Medicine
Program:
Authors:
Session: Session 3: 11:00 am – 11: 50 am
Poster: 94
Abstract
Does the CDKN2B gene upregulate IPF by expressing senescence-associated secretory phenotype (SASP) when the gene is isolated against IPF? It was found that when the CDKN2B gene was silenced, there was an increased differentiation of fibroblasts. Also observed was the lack of CDKN2B gene secreted and mediated in SASPs. METHODS: Materials were collected, RNA precipitation and washes were practiced, and slides were made. After, cells were counted, plated, and grown. This method includes working under the biosafety hood via vacuuming, pipetting, and centrifuging mixtures into RNA phases. Findings were analyzed by comparing the differences in gene expression for the two treatment groups: Control siRNA and CDKN2B siRNA, with a subtreatment among the groups stimulated with platelet-derived growth factor (PDGF) and fibroblast growth factor (FGF). The cells were again silenced with siRNA, and senescence was induced with hydrogen peroxide. RESULTS: We received enough samples to determine the silencing effects of CDKN2B in IPF. Initially, low cell counts were observed when reaching RNA analysis, but after restructuring the concentration necessary for cDNA conversion, this issue was mitigated. The slides were stained with ß-galactosidase and the supernatant was collected to observe cytokine protein expression. We predict that when CDKN2B is silenced, SASP expression is also downregulated, which promotes a higher level of senescence within IPF cell types. CONCLUSIONS: Silencing the CDKN2B gene causes an upregulation in myofibroblast differentiation, driving IPF. The data indicates there is evidence to prove a difference between silenced and expressed genes, even when stimulated PDGF and FGF and measured for senescence genes. These results suggest that more work needs to be done in this area, possibly providing insight that can be applied to the larger goal of IPF rehabilitation. Funded by: NIH grant F057376
